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Lorenzo, M. G.

Publications and source records attributed to Lorenzo, M. G..

7 recordsLinked to original sources

Cryo-EM structures of insect IRs reveal the structural adaptations of a glutamate receptor ancestor for environmental sensing

Ionotropic receptors (IRs) are an ancient, massively expanded family of sensory ion channels that mediate chemical, thermal, and hygrosensory detection across invertebrates. Derived from synaptic ionotropic glutamate receptors (iGluRs), IRs achieve this diverse tuning by assembling conserved co-receptor subunits with highly divergent tuning subunits into obligate heteromers. To understand how this architecture arose from a glutamate receptor ancestor we report cryo-EM structures of three insect IR complexes spanning the major functional subfamilies: IR8a/IR75b, IR25a/IR76b, and IR25a/IR93a/IR40a. All IR complexes retain the tiered iGluR architecture but adopt a distinctive splayed, non-swapped architecture that our phylogenetic reconstructions relate to a sister branch of glutamate delta (GluD) receptors. Our structures reveal a novel, co-receptor-specific helical beam feature that dictates stoichiometry by interlocking the co-receptors and separating the tuning subunits, at the cost of abolishing ligand binding in co-receptors. Comparison of IR8a- and IR25a- containing complexes further illuminates the structural basis of subfamily specificity yielding a predictive code of co-receptor assembly. Lastly, we identify IR-specific coupling mechanisms between the gating machineries of tuning and co-receptor subunits, offering a model for the cooperative gating of IRs. These analyses illuminate how an ancient synaptic receptor was architecturally remodeled into a multimodal family of sensory receptors that drive critical behaviors across invertebrates.

neuroscience↗

Global characterization of gene expression in the brain of starved immature R. prolixus

Background: Rhodnius prolixus is a vector of Chagas disease and has become a model organism to study physiology, behavior, and pathogen interaction. The publication of its genome allowed initiating a process of comparative characterization of the gene expression profiles of diverse organs exposed to varying conditions. Brain processes control the expression of behavior and, as such, mediate immediate adjustment to a changing environment, allowing organisms to maximize their chances to survive and reproduce. The expression of fundamental behavioral processes like feeding requires a fine control in triatomines because they obtain their blood meals from potential predators. Therefore, the characterization of gene expression profiles of key components modulating behavior in brain processes, like those of neuropeptide precursors and their receptors, seems fundamental. Here we study global gene expression profiles in the brain of starved R. prolixus fifth instar nymphs by means of RNASeq sequencing. Results: The expression of neuromodulatory genes such as those of precursors of neuropeptides, neurohormones, and their receptors; as well as the enzymes involved in the biosynthesis and processing of neuropeptides and biogenic amines were fully characterized. Other important gene targets such as neurotransmitter receptors, nuclear receptors, clock genes, sensory receptors, and takeouts were identified and their gene expression analyzed. Conclusion: We propose that the set of neuromodulation-related genes highly expressed in the brain of starved R. prolixus nymphs deserves functional characterization to allow the subsequent development of tools targeting them for bug control. As the brain is a complex structure that presents functionally-specialized areas, future studies should focus on characterizing gene expression profiles in target areas, e.g. mushroom bodies, to complement our current knowledge.

molecular biology↗

Breeding site microbiota and oviposition: do female Aedes aegypti shape bacterial communities?

Aedes aegypti, the main vector of multiple arboviruses, is highly associated with human dwellings. Females exhibit an opportunistic oviposition behavior, seldomly laying eggs on natural containers, but rather distributing them among human-generated breeding sites. Bacterial communities associated with such sites, as well as the compositional shifts they undergo through the development of larval stages, have been described. Some bacteria can play a direct role in supporting the success of mosquito development. Additionally, exposure to different bacteria during larval phases can have an impact on life-history traits. Whether the larvae acquire symbionts from aquatic niches, or just require bacteria as food, is still debated. Based on these facts, we hypothesized that female Ae. aegypti shape the bacterial communities of breeding sites during oviposition as a form of niche construction to favor offspring fitness. Our study presents a series of experiments to address whether gravid females modify bacterial consortia present in larval habitats. For this, we first verified if females can mechanically transfer bacteria into culture media. As evidence of mechanical transmission was obtained, we then elaborated an experimental scheme to dissect effects from factors related to the act of oviposition and mosquito-egg-water interactions. The DNA samples obtained from breeding site water belonging to five treatments were subjected to amplicon-oriented sequencing to infer their bacterial community structure. Microbial ecology analyses revealed significant differences between treatments in terms of diversity. Particularly, between-treatment shifts in abundance profiles were detected, also showing that females induce a significant decrease in alpha diversity through oviposition. In addition, indicator species analysis pinpointed bacterial taxa with significant predicting values and fidelity coefficients for the samples in which single females laid eggs. Furthermore, we provide evidence regarding how one of these indicator taxa, Elizabethkingia, exerts a positive effect upon the development and fitness of mosquito larvae, thus suggesting that the developmental niche construction hypothesis may hold in this model.

ecology↗

Changes in antennal gene expression underlying sensory system maturation in Rhodnius prolixus

BackgroundTriatomine bugs are the blood feeding insect vectors transmitting Chagas disease to humans, a neglected tropical disease that affects over 8 million people, mainly in Latin America. The behavioral responses to host cues and bug signals in Rhodnius prolixus are state dependent, i.e., they vary as a function of post-ecdysis age. At the molecular level, these changes in behavior are probably due to a modulation of peripheral and central processes. In the present study, we report a significant modulation of the expression of a large set of sensory-related genes. Results were generated by means of antennal transcriptomes of 5th instar larvae along the first week (days 0, 2, 4, 6 and 8) after ecdysis sequenced using the Illumina platform. ResultsAge induced significant changes in transcript abundance were established in more than 6,120 genes (54,7 % of 11,186 genes expressed) in the R. prolixus antenna. This was especially true between the first two days after ecdysis when more than 2,500 genes had their expression significantly altered. In contrast, expression profiles were almost identical between day 6 and 8, with only a few genes showing significant modulation of their expression. A total of 86 sensory receptors, odorant carriers and odorant degrading enzymes were significantly modulated across age points and clustered into three distinct expression profiles. ConclusionsThe set of sensory genes whose expression increased with age (profile 3) may include candidates underlying the increased responsiveness to host cues shown by R. prolixus during the first days after molting. For the first time, we describe the maturation process undergone at the molecular level by the peripheral sensory system is described in an hemimetabolous insect.

molecular biology↗

Evolution of the insect PPK gene family

Insect Pickpocket (PPK) receptors mediate the detection of stimuli of diverse sensory modalities, therefore having a relevant role for environmental sounding. Notwithstanding their relevance, studies on their evolution are scarce. We have analyzed the genomes of 26 species belonging to 8 insect orders (Blattodea, Orthoptera, Hemiptera, Phthiraptera, Hymenoptera, Lepidoptera, Coleoptera, and Diptera) to identify their PPK repertoires and study the evolution of this gene family. PPKs were detected in all genomes analyzed, with a total of 578 genes identified that distributed in 7 subfamilies. Our phylogenetic analysis allowed clarifying that the ppk17 gene appears to be the most divergent family member, composing a new group designed as subfamily VII. According to our analysis, PPKs evolved under a birth-and-death model that generated lineage-specific expansions usually located in clusters and the effect of strong purifying selection was seen for several orthogroups. Subfamily V was the largest one, presenting half of all PPKs studied, including a mosquito-specific expansion that can be considered a new target for pest control. Consistently with their sensory role, PPKs present a high gene turnover that generated considerable variation in the size of insect repertoires: Musca domestica (59), Blattella germanica (41), Culex quinquefasciatus (48), and Aedes albopictus (51) presented the largest PPK repertoires, while Pediculus humanus (only ppk17), bees and ants (6-9) had the smallest ones. The expansions identified in M. domestica and Bl. germanica also show promise as specific targets for controlling these nuisance insects. Our phylogenetic analysis revealed a subset of prevalent PPKs across insect genomes, suggesting a very conserved function that resembles the case of antennal ionotropic receptors. Finally, we identified new highly conserved residues in the second transmembrane domain that may be key for receptor function. Besides, more than a hundred PPK sequences presented calmodulin binding motifs, suggesting that at least some members of this family may amplify sensory responses as previously proposed for D. melanogaster ppk25. Overall, our study is a first attempt to characterize the evolutionary history of this family of sensory receptors, revealing relevant unknown features and clade-specific expansions.

genomics↗

Salty surfaces deter feeding in a blood-sucking disease vector

Salts are essential nutrients required for many physiological processes, and deficient or excessive salt results in adverse health problems. Taste is the ultimate sensory modality involved in resource quality assessment, resulting in acceptance or rejection. Here, we show that detection of high-salt substrates by a salt-sensitive antennal gustatory receptor neuron, S1-GRN, results in feeding avoidance in the hematophagous bug Rhodnius prolixus. Knock-down of two antennal-expressed amiloride-sensitive pickpocket channel receptors (PPKs; RproPPK014276 and RproPPK28) using RNA interference, prevents avoidance of bugs to high-salt substrates. Tracing antennal GRNs to the central nervous system reveals the antennal lobes as a gustatory processing center. The identification of the gustatory basis of high-salt detection in a blood feeder provides novel targets to prevent biting and feeding, as well as to promote substrate avoidance in a relevant disease vector. Significance StatementDetection of aversive gustatory stimuli induces avoidance responses in animals. Avoidance acquires particular interest if it reduces the biting rates of blood-feeding insects of medical relevance. Here we describe the molecular and physiological basis of high-salt detection in the blood-sucking disease vector Rhodnius prolixus. We show that detection of high-salt substrates through two PPK receptors expressed in an antennal gustatory receptor neuron produces feeding avoidance. Understanding these gustatory-driven aversive responses allows the hitherto overlooked use of gustatory molecules as a complement to known olfactory repellents.

physiology↗

Transcriptomics supports local sensory regulation in the antennae of the kissing bug Rhodnius prolixus

Rhodnius prolixus has become a model for revealing the molecular bases of insect sensory biology due to the publication of its genome, its well characterized behavioural repertoire and the advent of NGS technologies. Gene expression modulation underlies behaviour-triggering processes at peripheral and central levels. Still, the regulation of sensory-related gene transcription in sensory organs is poorly understood. Here we study the genetic bases of plasticity in antennal sensory function, using R. prolixus as an insect model. Antennal expression of neuromodulatory genes such as those coding for neuropeptides, neurohormones and their receptors was characterized by means of RNA-Seq. New nuclear receptor and takeout gene sequences were identified for this species, as well as those of enzymes involved in the biosynthesis and processing of neuropeptides and biogenic amines. We report a broad repertoire of neuromodulatory and endocrine genes expressed in antennae and suggest that they modulate sensory neuron function locally. Diverse neuropeptide-coding genes showed consistent expression in the antennae of all stages studied. Future studies should characterize the contribution of these modulatory components acting over antennal sensory processes to assess the relative contribution of peripheral and central regulatory systems on the plastic expression of insect behaviour.

neuroscience↗